8-hydroxy-11-desoxycorticosterones



8-HYDROXY-1l-DESOXYCORTICOSTERONES Herbert C. Murray, Hickory Corners,and Durey H. Peterson, Kalamazoo Township, Kalamazoo County, Mich.,assignors to The Upjohn Company, Kalamazoo, Mich, a corporation ofMichigan No Drawing. Application October 8, 1954,

Serial No. 461,284

7 Claims. (Cl. 260397.45)

The present invention relates to steroid and is more particularlyconcerned with 8-hydroxy-1l-desoxycorticosterone and the 21-acyloxyesters thereof represented by the following formula:

CHzOR wherein R is selected from the group consisting of hydrogen andthe acyl radical of a hydrocarbon carboxylic acid containing from one toeight carbon atoms, inclusive.

The S-hydroxy-ll-desoxycorticosterone is prepared by exposingll-desoxycorticosterone or any of the 21-esters ofll-desoxycorticosterone to the oxygenating action of a fungus of theorder mucorales, preferably of the family Mucoraceae and the genus Mucorin a suitable medium and with aeration as shown in detail in theexamples.

In the preparation of the ester of the 8-hydroxy-11-desoxycorticosterone, the starting 8-hydroxy-l1-desoxycorticosterone isadmixed with an acylating agent such as, for example, ketene, a keteneof a selected acid, an acid, acid chloride, or acid bromide, or an acidanhydride, or other known acylating agent, usually in a solvent such as,for example, pyridine or the like, or in an inert solvent, includingsolvents like benzene, toluene, ether, and the like, and heated at atemperature between about zero degrees centigrade and the boiling pointof the reaction mixture, usually about room temperature, for a periodbetween about one half hour and about 96 hours. The time of reaction issomewhat dependent upon the temperature at which the reaction is carriedout, the acylating agent, and the ratio of reactants. The reactionmixture is quenched with ice or cold water and the product is collectedin an organic solvent which is thereafter washed with a mildly basicsolution and water to obtain a solution of the product which isessentially neutral. In some instances, the product may be crystallizedfrom the reaction mixture, in which event it may be advantageous toseparate the product by filtration or other means, to wash it withwater, and thereafter to purify it by conventional means, such as, forexample, by re-, crystallization from a suitable solvent or bychromatographic purification, as deemed necessary.

It is an object of the present invention to provide8-hydroxy-ll-desoxycorticosterone and the ZI-hydrocarbon carboxylic acidesters thereof. Other objects will be apparent to those skilled in theart to which this invention pertains.

Thecompounds of the present invention demonstrate pronounced anesthetic,salt retention, bacteriostatic and nited States Patent 0 fungistaticactivity. The compounds of the instant invention are also important asintermediates in the production of active steroids. For example,vigorous oxidation of 8-hydroxy-1l-desoxycorticosterone with chromic an-5 hydride in actic acid gives known 8-hydroxy-4-androstene-3,l7-dione.The instant compound may also be used for the preparation of the known21-hydroxy-l4- pregnene-3,20-dione. This preparation may be carried outby first reducing 8-hydroxy-ll-desoxycorticosterone with hydrogen in thepresence of a palladium catalyst under alkaline conditions to give8,2l-dihydroxypregnane- 3,20-dione which is dehydrated with methanolichydrochloric acid to give 2l-hydroxy-8(9) -pregnene-3,20-dione which canbe isomerized to ZI-hydroxy-l4-pregnene-3,20- dione [Reichstein et all,Helv. Chim. Acta 30, 1508 (1947)] with the aid of perchloric acid indioxane solution.

The starting material of the present invention is 11"-desoxycorticosterone and the 2l-esters thereof. The 21- acetate ofll-desoxycorticosterone is an already known compound [Steiger et al.,Helv. Chim. Acta 20, 1164 (1937)]. While other esters may be prepared bythe usual methods of esterification, such as illustrated herein, thereis, however, no particular advantage in using the more expensive esterssince during the S-hydroxylation the ester group is split oil from theoriginal ll-desoxycorticosterone by the enzymatic activity of thefungus.

The following examples illustrate the process and products of thisinvention, but the invention is not to be construed as limited thereto.

EXAMPLE l. 8 HYDROXY ll DESOXYCORTI- COSTERONE sterilized medium thusprepared were inoculated with Mucor parasiticus (American Type CultureCollection No. 6476), and incubated for 24 hours at a temperature of 28degrees Centigrade using a rate of aeration and stirring such that theoxygen uptake was 6.3 to- 7 millimoles per hour per liter of NazSOaaccording to the method of Cooper, Fernstrom and Miller, Ind. Eng. Chem.36, 504 (1944). A solution of 1.1 grams of desoxycorticosterone acetatedissolved in 250 milliliters of acetone was divided into four parts andone part was added to each of the four bottles containing a 24 hourgrowth of Mucor parasiticus. After an additional 24 hour period ofincubation for two of the bottles the beer and mycelium were extracted.The other two bottles were incubated for 48 hours and then extracted.The mycelium was filtered, washed twice, each time with a volume ofacetone approximately equal to the volume of the mycelium and extractedtwice, each time with a volume of methylene chloride, approximatelyequal to the volume of the mycelium. The acetone and methylene chlorideextracts including solvent were added to the beer filtrate. The mixedextracts and beer'filtrate were extracted successively with two one-halfby volume portions of methylene chloride and with two one-fourth byvolume portions of methylene chloride. The combined methylene chlorideextracts were washed with two onetenth by volume portions of a twopercent aqueous solution of sodium bicarbonate and then with twoone-tenth by volume portions of water. After drying the methylenechloride extracts with about three to five grams of anhydrous sodiumsulfate per liter of solvent and filtering, the solvent was removed bydistillation. The residue was dissolved in a minimum of methylenechloride,

'3 2 filtered and the solvents then evaporated. The resulting residueswere then combined to give 27.35 grams of solids which were dissolved in500 milliliters of ethylene dichloride and chromatographed over 900grams of Florisil synthetic magnesium silicate. Fractions of 1400milliliters were collected as indicated in Table I. g

Table I Fraction Solvent Eluate, grams 1 Ethylene dichloride 3, 508 2 do1.319 3... Ethylene dichloride-acetone 2511.. 1. 596 4--. do 0.123 5.Ethylene dichloride-acetone 15:1. 0. 212 6- d 1.237 7. Ethylenedichloride-acetone 12: 2. 328 8- ..do 1.019 9- 0.924 10-. 1.045 11 l1.000 '12" 0.803 13 0.928 14 0.747 15 0.656 16.; 0.966 17;. 0.817 182.095 19-. 0.769 20. Acetone 0. 678

Eluate fractions 18 and 19 were combined to give 2.665 grams of materialwhich was dissolved in 150 milliliters of ethylene dichloride andrechromatographed over 150 grams of Florisil. Fractions of 250milliliters were collected as shown in Table II.

Table II Fraction Solvent Eluate,

milligrams Ethylene dichloride Ethglene dichloride-acetone 10: 1- 0 H...o Acetone 1 ,Eluate fractions 10 and 11, found to be crystalline afterevaporation of the solvent, were combined, dissolved in ten millilitersof acetone which was then concentrated and gradually diluted withpetroleum ether to effect crystallization. After two recrystallizations,134.5 milligrams of crystals, melting point 180 to 183 degreescentigrade, were obtained. A small sample was recrystallized once morefrom acetone-Skellysolve B (a mixture of hexanes) to give the pure8-hydroxy-1l-desoxycorticosterone, M. P. 182.5-184". Ultravioletspectrum 243mp,E 14, 300, [04 167 4 (ATCC 8686, and 8992), in a nutrientmedium with aeration. Isolation of the thus obtained8-hydroxy-lldesoxycorticosterone is generally carried out bychromatography. The preferred solvent in the chromatographic proceduresare methylene dichloride, ethylene dichloride, acetone and mixturesthereof.

EXAMPLE 2.8-HYDROXY-1l-DESOXYCORTI- COSTERONE Zl-ACETATE A solution of44.5 milligrams of S-hydroxy-ll-desoxycorticosterone in threemilliliters of a solution of pyridine and acetic anhydride in a ratio ofone to one was allowed to stand for a period of 48 hours at roomtemperature. Thereafter the reaction mixture was poured into ice waterand the thus-obtained mixture was extracted three times with methylenedichloride. The extracts were washed once With five percent hydrochloricacid, then with five percent sodium carbonate and twice with Water. Upondrying of the methylene solution over anhydrous sodium sulfate andconcentrating, 52.2 milligrams of a crystalline residue Were obtainedWhich were recrystallized twice from two milliliters of acetone to whichSkellysolve B was added dropwise. In this manner 37.6 milligrams of8-hydroxy-ll-desoxycorticosterone Zl-acetate was obtained, melting point212 to 215 degrees centigrade, rotation [M plus 177 degrees at aconcentration of 0.834 in chloroform.

Analysis.Calculated for CzsHzzOs: C, 71.10; H, 8.30. Found: C, 71.14; H,8.08.

EXAMPLE 3.8-HYDROXY-l l-DESOXYCORTI- COSTERONE ZI-PROPIONATE In themanner given in Example 2, 8-hydroxy-lldesoxycorticosterone was reactedwith propionic anhydride ill pyridine solution to yield8-hydroxy-ll-desoxycorticosterone 21-propionate.

EXAMPLE 4.8-HYDROXY-l l-DESOXYCORTI- COSTERONE ZI-BUTYRATE In the mannergiven in Example 2, 8-hydroxy-lldesoxycorticosterone was reacted withbutyric anhydride in pyridine solution to yieldS-hydroxy-ll-desoxycorticosterone 21-butyrate.

EXAMPLE 5.--8-HYDROXY-1 l-DESOXYCORTI- COSTERONE ZI-VALERATE In themanner given in Example 2, 8-hydroxy-lldesoxycorticosterone was reactedwith valeric anhydride in pyridine solution to yield S-hydroxy-ll-desoxycorticosterone 2 l-valerate.

EXAMPLE 6.-8-HYDROXY-1lDESOXYCORTI- COSTERONE 2 l-HEXANOATE In themanner given in Example 2, 8-hydroxy-lldesoxycorticosterone was reactedwith hexanoic anhydride in pyridine solution to yield8-hydroxy-ll-desoxycorticosterone 2l-hexanoate.

EXAMPLE 7.8-HYDROXY -1 l-DESOXYCORTI- COSTERONE ZI-BENZOATE In themanner given in Example 2, 8-hydroxy-lldesoxycorticosterone was reactedwith a solution of benzoyl chloride in pyridine to yield8-hydroxy-1l-desoxycorticosterone 21-benzoate.

, 8.8-HYDROXY-l l-DESOXYCORTI- COSTERONE Zl-TRIMETHYLACETATE EXAMPLE 9.s HYDROXY 11 DESOXYCORTI- COSTERONE 2l-(fl-CYCLOPENTYLPROPIONATE) "In'the manner given in Example 2, 8-hydroxy-1ldesoxycorticosterone wasreacted with a solution of cyclopentylpropionyl chloride in pyridine toyield 8-hydroxy- 1 l-desoxycorticosterone 21-(IS-cyclopentylpropionate)EXAMPLE 10.--8-HYDROXY-1 l-DESOXYCORTI- COSTERONE ZI-SALICYLATE cludeone to eight carbon atom carboxylic acid acyloxy 2 esters of saturatedor unsaturated, aliphatic, carbocyclic or cycloaliphatic, aryl,arylalkyl, alkaryl, mono-, dior polycarboxylic acid esters such as the8-hydroxy-11-desoxycorticosterone 21-fo1'mate, isovalerate, heptanoate,octanoate, cyclohexylformate, toluate, anisate, gallate, phenylacetate,cinnamate, vinylacetate, dimethylacetate, hemisuccinate, hemitartrate,dihydrogencitrate, glutarate, maleate, malonate, and the like.

It is to be understood that the invention is not to be wherein R isselected from the group consisting of hydrogen and the acyl group of ahydrocarbon carboxylic acid containing from one to eight carbon atoms,inclusive.

2. 8-Hydroxy-1l-desoxycorticosterone. 5 3.8-Hydroxy-11-desoxycorticosterone 21-esters of the formula:

(llHaoR C=O 10 CH:

wherein R is the acyl radical of a hydrocarbon carboxylic 0 acidcontaining from one to eight carbon atoms, inclusive.

4. 8-HydroXy-1l-desoxycorticosterone 21-acetate. 5.8-HydroXy-1l-desoxycorticosterone 21-propionate. 6.8-Hydroxy-1l-desoxycorticosterone ZI-(fl-cyclopentylpropionate) 25 7.8-Hydroxy-1l-desoxycorticosterone 21-benzoate.

References Cited in the file of this patent UNITED STATES PATENTS2,602,769 Murray et a1 July 8, 1952 limited to the exact details orexact compounds shown and described, as obvious modifications andequivalents will be apparent to one skilled in the art, and theinvention is therefore to be limited only by the scope of the appendedclaims.

We claim:

1. An S-hydroxy-ll-desoxycorticosterone of the formula:

CHzOR

1. AN 8-HYDROXY-11-DESOXYCORTICOSTERONE OF THE FORMULA: